Act1分子对骨髓瘤细胞增殖的负调节作用研究

目的:探讨调节因子Act1对骨髓瘤细胞中BAFF(B细胞活化因子)通路的调节作用,进而了解其对骨髓瘤细胞的增殖存活所产生的作用.方法:利用荧光定量PCR的方法测定20例多发性骨髓瘤(MM)病人及20例正常对照者的外周血中的BAFF分子和Act1分子的表达水平;通过小干扰RNA(siRNA)和基因过表达技术构建缺乏和过表达Act1的U266骨髓瘤细胞,利用免疫印迹(Western blot,WB)法测定骨髓瘤细胞在BAFF分子刺激下的NF-κB信号通路活化情况,即磷酸化的IκB(p-IκB)的表达水平.结果:MM病人外周血BAFF和Act1分子的表达水平较对照组均增高(P<0.01),二者之间亦存在相关性;缺乏Act1分子的骨髓瘤细胞IκB磷酸化水平较高,而对照组的细胞IκB磷酸化程度较低,趋势不明显,过表达Act1分子的细胞基本没有磷酸化的发生.结论:Act1分子在多发性骨髓瘤的发生发展中发挥重要作用,其作用是通过对BAFF分子刺激下NF-κB信号通路的负调节作用而发挥的,使其可能成为抑制骨髓瘤细胞增殖的作用靶点.

Negative regulation of Act1 to the myeloma cell proliferation

Objective:To investigate how the BAFF signal is affected by Act1,and to find the function of Act1 to the proliferation and living in myeloma cells.Methods:The expression of BAFF and Act1 genes in MM patients were performed with RTQ-PCR,the siRNA of Act1 and the synthesized Act1 gene were transferred to U266 cells respectively to cut down or overexpress the Act1 protein,then the western blot was performed on the p-IκB by stimulating with BAFF in different times.Results:The mRNA of BAFF and Act1 were all increased in MM patients compared to the healthy persons from our experiments.The p-IκB increased more highly in the cells transferred with siRNA than in the untreated cells,but the cells overexpressed the Act1 changed very mildly.Conclusion:It was important of BAFF and Act1 in the generation and development of MM,and the result proved the negative regulation of Act1 to BAFF signal passway in MM cells.The Act1 could be a target spot for the treatment of MM.